Raw grain and feed compositions containing 1,3-aliphatic diols and/or their esters

ABSTRACT

Raw grain and feed compositions of great resistance to the attack of molds, bacteria and yeast are prepared by utilizing with said substances an aliphatic 1,3-diol and/or their esters.

United States Patent [1 1 Frankenfeld et al.

[22] Filed: Jan. 27, 1971 [21] Appl. No.: 110,247

[52] US. Cl 426/151, 426/208, 426/210 [51] Int. Cl. A23k 3/02, A23b 9/00[58] Field of Search 99/8, 17, 150 R,

[56] References Cited UNITED STATES PATENTS 1,802,083 4/1931 Legendre99/153 1 51 Nov. 20, 1973 2,437,150 3/1948 Berg 99/83 2,651,572 9/1953Bickoff 99/8 3,145,109 8/1964 Howard 99/123 X 3,373,044 3/1968 Thompsonet a1... 99/123 X 3,479,189 11/1969 Vrang et al. 99/91 X 3,485,63912/1969 Knightly 99/150 X 3,667,964 6/1972 Frankenfeld et a1.... 99/150X 3,667,965 6/ 1972 Fiankenfeld et al 99/150 X Primary Examiner-NormanYudkoff Assistant Examiner-Kenneth P. Van Wyck Att0mey-Manahan andWohlers and W. O. Heilman 57 ABSTRACT Raw grain and feed compositions ofgreat resistance to the attack of molds, bacteria and yeast are preparedby utilizing with said substances an aliphatic 1,3-diol and/or theiresters.

11 Claims, No Drawings 1 RAW GRAIN AND FEED COMPOSITIONS CONTAININGl,3-ALIPHATIC DIOLS AND/OR The present invention is concerned with rawgrain and feed compositions which are stable in storage and duringshipment against the attack of molds, yeasts and bacteria.'Thisstability and resistance to attack is secured by utilizing inconjunction with these materials arelatively small amount of al,3-aliphatic diol and/or their monoor diesters.

A very great problem in the storage and transportation.of raw grains andfood compositions is that they are subject to attack of various fungi,molds, yeasts and bacteria which, in many instances, render these foodmaterials poisonous both to animals and to humans. Large quantities ofgrain are grown to feed all classes of livestock, and microbiologicaldeterioration of the food products in storage is a very serious problem.The molds and bacteria which can develop in stored grain not onlyconsume its more valuable nutritive components but as heretoforementioned are also a hazard to the health of both humans and animals.

Various techniques had been developed in order to at least partiallysolve this problem. One technique is to reduce the moisture content, forexample below about 14 percent. Another technique is to use an additivesuch as propionic acid. Other techniques are to restrict the growth ofthe microorganism by depriving them of oxygen or by chilling.Particularly undesirable and harmful microorganisms are, for example,Bacillus mesentericus, Aspergillus, Penicillium and Salmonellamontevideo. These microorganisms particularly attack the grains such aswheat, rice, oats, rye, beans, maize and hay. Many of thesemicroorganisms produce mycotoxins which are harmful to humans andanimals. A particularly dangerous mycotoxin producer is AspergiIlusflavus.

Thus, the present invention is broadly concerned with the control ofmicroorganisms which attack grains and other forage crops. Some of themicroorganisms among many others which attack grain and relatedsubstances are as follows:

Achromobacter spec. Monilia spec. Alremaria flavu: Mucor Aspergillusflavus Oospora Aspergillus fumigalus Penicillium digilarum Penicilliumglaucum Penicillium notalum Pezisella lylhri Pseudomonas aeruginosaPseudomonas nanliquefaciens Pullularia pullulans Rhizopul nigricansRhodammla .rpec Salmonella Saccharomyres cerevisiae Saccharamycesellipsoideus Sparendomema epizoum Sireplobacleriam platarum lipalyricaTarulopsir caroliniana Tarulopsis homii Trichosporon variable Williaanomala Zygasaccharam yces halomenbrani:

Thus, the present invention is concerned with feed compositions, rawgrain compositions and the like which contain intimately mixed therewitha relatively small amount of a l,3-aliphatic diol and/or their monoordiesters. The grain granules are, in effect, coated with a fine film ofthe diol or diol ester which may be applied by any suitable techniquesuch as by mass mixing or by spraying.

The amount of diol or diol ester utilized is in the range from about0.05 to 2.0 percent by weight, preferably in the range from about 0.1 to0.5 percent by weight based upon the weight of the material beingpreserved against attack.

The 1,3-diol contains from about six to 15 carbon atoms in the diolchain, preferably from seven to nine carbon atoms in the diol chain.Very desirable diols for utilization in the present invention arel,3-heptanediol, l,3-octanediol and 1,3-nonanediol. If a monoor diesterbe utilized, the number of carbon atoms in the diol chain is in therange from about three to 15, preferably in the range from four to 10.The number of carbon atoms in the ester group may vary in the range fromabout two to 15, preferably from about three to 10. Particularlydesirable diol esters are l,3-octanediol-lmonopropionate and l,3-butanediol-1- monooctanoate.

The particular preferred diols of the present invention are l,3-diolsand correspond to the following i al sys alssms.-.

RCHCHICHI where R represents an n-alkyl group containing three to 12carbon atoms. Thus, the preferred 1,3-diols must contain a minimum ofsix carbon atoms in the chain (R 3). Diols with fewer carbon atoms arenot as effective preservatives. The preferred diols contain from sevento nine carbon atoms in the chain (R 4 to 6).

The preferred monoesters of the present invention correspond to thefollowing chemical formulas:

RCHCHQCH} RCHCHrCI-I,

H C R; C R; H

where R an n-alkyl chain of zero to 12 carbon atoms and R, is an n-alkylchain containing from one to 17 carbon atoms. Thus, the esters maycontain from three to 15 carbon atoms in the diol portion of themolecule. The preferred monoesters are those which contain four to 10carbon atoms in the diol portion (R 1 to 7) of the molecule and fromthree to 10 carbon atoms (R 2 m9) in the ester portion of the molecule.Very desirable monoesters are l,3-butanediol-1-monooctanoate and1,3-octanediol-l-monopropionate. These monoesters may exist in twodifferent forms. Form A is the lmonoester form. Form B is the3-monoester forrn.

Still another ester form which is useful for certain applications is thediester C.

RCHGHiCHI where R an n-alkyl chain of zero to 12 carbon atoms and R, isan n-alkyl chain of one to 17 carbon atoms. The preferred diesterscontain from about four to eight carbon atoms in he diol portion (R l to5) and from three to -10 carbon atoms (R,.=-2.to 9') in the esterportions of the molecule. Although the diesters are generally not asactive as the monoesters, they are useful in cases where their physicalproperties (e.g., their increased lipophilic character) are especiallydesirable.

It is essential, for both the l,3-diols and their esters that themolecules be linear and that the hydroxy and- /or carbalkoxy (ester)groups be attached to the first and third carbon atoms. it is thisl,3-difunctionality which renders these compounds especially usefulbecause of their inherent safety for use around humans and animals.

Other suitable diols for use in the present invention arel,3-hexanediol, l,3-decanediol and 1,3- undecanediol.

The diol and diol esters of the present invention may be prepared by anysuitable technique such as by the Reformatsky reaction followed byreduction, or by means of the Prins reaction of formaldehyde and theappropriate olefin.

In order to further illustrate the invention, various tests were carriedout, the results of which are described in the following examples andtables of data. In Table l are summarized the results of toxicitymeasurements and nutritional evaluations for a variety of dihydroxycompounds including the l,3-diols. These studies were conduced withrats.

The observed metabolic energy values shown in Table l were obtained byfeeding test groups, of fiverats each, various amounts of several highenergy supplements including the 1,3-diols. The basal diets in each casecontained sufficient protein, salts, vitamins and minerals to supportnormal growth. However, the basal diets were deficient in energy(calories). This deficit was, in part, overcome by adding varyingamounts of the polyols or of natural energy sources of known caloricdensities such as lard, sucrose, or glucose. Curves were drawn byplotting the average change in body weight of test animals against theamount of high energy supplement tested and straight lines wereobtained. The slopes of the lines are measures of the energy values ofthe test compounds. The values given in Table l were obtained bymeasuring the slopes of the lines of the test compounds and comparingthem with the slopes of the lines obtained with the standards (lard,glucose or sucrose whose caloric densities are 9.3 Kcal/g, 3.8 Kcal/gand 4.1 Kcal/g, respectively). The observed metabolic energy" valueswere calculated according to the formula:

I W observed metabolic energy of a material is a measure of itsnutritional value. The higher the energy the greater its value. It isclear from the results given in Table I that 1,3-diols are verydesirable components from a safety standpoint. Compounds which do notposses the 1,3-dihydroxy structure are toxic and, therefore, not usefulpreservatives where contact with humans or animals is likely. V 5 ED,values are a common measure of the toxicity of a compound. These LDvalues represent the lethal dose for a 50 percent kill of the animalstested per unit weight of the animals. The higher the LD value, thelower the toxicity. The data in Table l establish that the1,3-configurations of polyalcohols are the least toxic. The LD valuesare significantly higher in compounds possessing this structuralfeature. The LD values 7 Oral Observed LDm metabolic Dlhydroxy (7 dayenergy Percent Compound compound gJkgJ 2 (KcnL/g.) utilized 31,3-butunedi0l 29 6.0 1 3-pentanediol 20 7. 8 1,6-pentanedio 2 NU1,3-hexanediol. 20 6. 6 1,5-hexanedioL 20 4 NU 2,5-hexanediol 2 4 NU1,6-hexanedi0l 5 4 NU 1,3-heptanediol 20 8.0 1,3-octanediol 20 5. 81,3-nonanediol 20 7. 5 1,3-decanediol 20 12 1,3-undecanediol- 20 1Single dose in rats.

1 LD5 =1ethal dose for 50% kill.

Determined by dividing observed metabolic energy (Kcal.) by theoreticalmetabolic energy (Kcal.).

4 N U=Not Utilized.

given in Table I were obtained by giving test animals graded singledoses of the test compounds orally and observing them for one week. Thenumber of deaths in each group was noted and the dose required for a 50percent kill taken as the LB value. In many cases, no

death occurred even at the 20 g/kg level (about as much as one can givea rat in one dose), hence the basis of the 20 values shown in the table.

The toxicity data with respect to the esters are given in the followingTable II. The LD values for some common food and feed preservatives arealso shown. All are significantly more toxic (lower LD values) thaneither the diols or the esters.

Table II TOXICITY DATA Preservative Oral LD (Rats) Diol esters 20 g/Kg(propionates and higher) Sorbic acid 10 g/Kg gSodium sorbate 6-7 g/KgPropionic acid 4 g/Kg (Sodium benzoatc 2-3 g/Kg .LD Flcthal dose for 50%kill.

"Source: Handbook of Toxicology, Vol. I, W.S. Specter, ed., \VADC Teclifiept. NB. 55-16, National Academy of Sciences, National ResearchCouncil ([955).

Source: H.F. Smyth et al., Am. lnd. Hyg. Assoc. J., 23, (1962) In orderto further illustrate this invention and, in

particular, to establish the superiority of the l,3-diols and esters asbacteriostats and mold inhibitors, the following microbiological testswere conducted.

EXAMPLE 1 Nutrient broth was used as the basal nutrient medium for thegrowth of all microorganisms tested. Five ml. of nutrient broth medium(Difco Co.) were placed in 18 mm X mm test tubes and the basal mediumsterilized with steam at 15 psi for 15 minutes. After cooling, asufficient amount of the various compounds were added to the basalmedium to give the concentrations used. Normally a final concentrationof 0.2, l and 2 percent were used.

After mixing the chemicals with nutrient broth, the tubes wereinoculated with the various test microorganisms. The test microorganismswere grown25 hours earlier in nutrient broth and 1 drop of the densemicrobial suspension was added to the tubes.

The tubes containing the chemicals and microorganisms were thenincubated at the optimal growth temperature reported for eachmicroorganism tested. Either 37 C. or 30 C. was used. Growth in controltubes, as well as those containing chemicals, was observed visually.After a suitable incubation period, a small aliquot of the testsolutions was streaked on an agar plate.

This was done in order to confirm the visual readings of the presence ofmicrobial growth. The results are shown in Table III and IV. The minimumeffective concentration is the lowest concentration of additive whicheffectively prevented growthunder the conditions of the test.

It is apparent from the above that the effectiveness of these materialsagainst a wide spectrum of bacteria is TABLE IV.-ACTION AGAINST FUNGIgrowth even at the 2 p ercentlevel. salmonellae alert Compound1,3-pcntanediol 1,3-heptanediol... 1,3-pentanediol-monopropionat1,3-octanediol-monopropionate l,3-butanediol-dipropionate.l,3-butanediol-monooctanoate. 1,B-pentanediol-monooctanoate Potassiumsorbate Calcium propionate Minimum inhibitory concentration (percent)against- P. requertii 6988 I Fusarium 10911 1 Norm-Symbols: +=No efiectat 2%. The lowest level tested in this experiment was 0.2%. =Not tested.1 All microorganism numbers, American Type Culture Collection.

established by the typical data shown in Table Ill. In he tests the testp und e e sneelsd 23. 12

In Table V some selected diols and esters are comwith. eta s e f sereteiqds iem BEOBLJRFL V.MINIMUM INHIBITORY CONCENTRATION (PERCENT)AGAINST TABLE ASPERGILLUS NIGER UNDER SPECIFIED CONDITIONS Dextroseadded, wt. Glycerol added, wt.

percentpercent- Nutrient broth 5 5 10 20 Potassium sorbato X X X X X X XPropylene glycol" X X X X X X X 1,3-butuucdiol XX XX XX XX XX XX XXCalcium propionotc. X 03% Olg OX) 0 0 0);,

- 1. 0 0. 5 1. 0 0. 5 0. 5 1. 0 1,3-butancdiol-monooctanoatc 0. 05 0. 050. 05 0. 05 0. 05 0. 05 O. 05 l ,8-octauediol-monopropionate 0. 05 0. 050. 05 0. 05 0. 05 O. 05 1,3-nonancdiol 0. 05 O O O O O 0 Potassiumsorbate 1. 0 X X X X X 0. 2 2. 0 1. 0 Calcium propionate X X X X X X X1-,3-heptancdiol 0.1 0. 2 0. 2' 0. 2 0. 2 0. 2 0. 2 0.5 0.5 0.5 0.5 0.5.5 1,3-butanediol-monooctanoate 0. 1 0. 1 0. 1 0. 1 0. 1 0. l 0. 1 0.10.1 0.1 0.1 0.1 0.1 0.1

1,3pctanediol-monopropionate NoTE.-X=Not effective at the highest leveltested: 2%. XX=Not efiective at the highest level tested: 5%.

O Not tested.

known commercial preservatives, potassium sorbate commercial moldinhibitors, in effectiveness against and calcium propionate, as to theirability to TABLE IIL-ACTION AGAINST BACTERIA Minimum inhibitoryconcentration (percent) against- Salmonella P. aerutyphi- Compoundginoau murium E. coli 1,3-heptanediol 1 0. 5 1 l 1,3-nonanediol 0. 5 0.2 O l,3-octanediol-l-monopropionate O 0. 2 2 1,3-butanedioldipropionateO 0. 2 1 Potassium sorbate 2 Calcium propionate Nora-Symbols: +=Noefl'ect at 2%. O=not tested.

- growth of various bacteria. It isapparent that several of variouscommon'molds. The lower the minimum effective concentration, the moreeffective the compound. It is apparent that several of the testcompounds are significantly better than the currently used preser- 0vatives.

EXAMPLE .2

In addition to these tests, some more definitive stud- .ies wereconducted to determine the effectiveness of certain diols and esters ininhibiting'the growth of two common molds under various culture'conditions. These tests-were carried out as described above except thatthe pH was varied by the use of suitable buffering agents and, in someexperiments, either dextrose or glycerol were added to demonstrate theeffectiveness of the diols and esters in different growth-media.

For purposes of comparison, several commercial preservatives were testedunder the same conditions. The results are shown in Tables V and VI. Itis apparent from these data that the 1,3-diols and esters are significantly'more effective than the commercial additives. Of especialimportance is the finding (Tables V and VI) that 1,3-heptanediol and theesters are highly active inhibiters 9f. W5 .HPFQFQQPYALGQQQEQ. reaelieitii 8 RESULTS Batch Additive Percent Mold Score 1 None (control) 21,3-Heptanediol 0.50 3 3 l ,3-Heptanediol 0.75 2 4 1,3-Heptanediol 1.00l

Thus, the present invention is concerned with feed and feed compositionswhich have excellent resistance to attack by molds, bacteria and yeastwhich contain as an ingredient thereof a l,3-diol and/or ester asdefined heretofore.

TABLE VL-MINIMUM INHIBITORY CONCENTRATION (PERCENT) AGAINST PENICILLIUMROQUEFORTII UNDER SPECIFIED CONDITIONS Dextrose added, wt. percentGlycerol added, wt. percent Nutrient broth 5 5 10 20 02 02 02 0.2 02 020.2 Pmassmm bate 1. 0 1. 0 1. 0 1. 0 1. 0 1. 0 1. 0 Calcium propionateXX XX X); XX O.2 0. 02 heptanedml O5 0. 5 0. 5 0. 5 0. 5 0. 5 0. 51,3-butanedioI-monooetan0ate. O. 05 0. 05 0. 05 O. 05 0. 05 0. 05 0. 051,3-oetanediol-monopropionate O. 05 0. 05 0. 05 0. O5 0. 05 0. 05 0. 05

- 02 02 02 02 02 02 0.2 Pnassum 50mm 1. 0 1. 0 1. 0 1. 0 1. 0 1. 0 1, 0Calcium proplonate XX Iglg XX X XX XX XX 0-2 0. 02 O.2 0.2 helmmnol 0.50.5 0.5 0.5 o.5 05 1,fl-butenediol-monooctanoate 0. l 0. l 0. l 0. 1 0.1 0. 1 0. 1 1,3-0etanediol-monoproplonate 0. 1 0. 1 0. 1 0. 1 0. 1 0. 10. 1

N o'rE.--XX=Not efieetive at the highest level tested: 2%.

The d ols and/or diol esters of the present invention TABLE VI" areparticularly desirable for use 1n feed compositions such as chicken feedcompositions. A suitable feed CALCULATED COMPOSITION mash is listed inTable Vll and a composition in Table W VIII. The diols and/or diolesters are included in the Percent feed at a level of 0.1 to 0.5percent, that is, 2 to 5 p i (N X 625) 230 pounds per I011. Arg inine 15r Lysine l.0l TABLE VII.FORMULA 1a-ALL MASH STARTING FEED 40 glethlomneystlne Pounds per Pounds per Tl-ypmphane 013 Ingredient 100 pounds tonGlycine more than L4 Ground ellow corn 25.00 500 lsoielfcine 1.16Pulverizg d oats 15. 00 300 Lemme 2.3l Wheat middlings, standard 15.00300 Phenylalanine 1.14 Soybean meal 2- 2 Th oni Q86 Corn gluten me 9.75195 Valine 8 Fish meal 8.50 170 Dried skim milk- 2. 00 so e 0.55Dehydrated alfalfa meal. 7. 60 160 Tyrosine 0.85 Dried dlstllierssolubles. 2. 50 Fat (ether extract) 4.3 Mineral mixture No. 1 1.30 26Crude fiber (L5 13,""55113561166153 3' 1; 3 Calcium ee ng 50 Phosphorus0.73 Total 100.00 2,000 Total salt (NaCl) 0,59 Pt, 0.85 Magnesium 0.23Parts per Million EXA 3 Manganese 62 iodine 6 Evaluation of l,3-Dlols asPreservatives for Poultry on more h a n 80 Feeds Vitamin A units perpound h 6000-9000 PROCEDURE Vitamin D A.O.A.C. (chick units per pound)270 To 100 g batches of FCA Chick Starter (see Formula Milligrams perPound la) were added 5 ml of water and various amounts of the additivesshown below. The batches were then pamlnhenic acid stored at roomtemperature for 72 hours and evaluated S 0 630 mom 1 for mold growth ona scale from l-5. Thiamine 26 5 Heavy mold g 5.85 PPM is supplied byMineral Mixture No. 1; only about 0.15 to 4 Moderate mold grgwth, 0.20PPM is supplied by the other ingredients.

3 Light growth. 2 Slight to questionable growth. I No growth.

Depends chiefly on the carotene content of the alfalfa meal.

The effectiveness of the diols of the present invention is illustratedin the following example.

What is claimed is: i i 1. Raw grain composition comprising raw grainhaving admixed therewith from 0.05 to 2.0 percent by weight based on theweight of grain present of an aliphatic 1,3-diol or an ester thereof,said diol character- I monopropionate.

4. Composition as defined by claim 1 wherein the amount of diol or diolester present in the rangefrorn about 0.1 to 0.5 pe rcent by weightbased upon the weight of the grain.

5. Composition as defined by claim 1 wherein said raw grain compriseswheat particles.

6. Composition as defined by claim 1 wherein said raw grain comprisesoats.

7. Composition as defined by claim 1 wherein said 'raw grain comprisesrye.

8. Composition as defined by claim 1 wherein said raw grain comprisesbeans.

9. Composition as defined by claim 1 wherein said raw grain comprisesmaize.

l0. Composition as defined by claim 1 wherein said raw grain compriseshay.

l1. Composition as defined by claim 1 wherein said raw grain comprisesrice.

UNITED STATES PATENT oFFicE 1 7 CERTIFICATE: OF CRECT IQN Dated biiovembr 20,1973

Iatent No. 3,775,518

inventoflsflohn W. Frankenfeld, Raam R. Mohan and Robert L. Squibb It iscertified that error appears in the above-identified patent and thatsaid Letters Patent are hereby corrected as shown below:

-- Assignee: I Esso Research and Engineering Comparry,

Signed and sealed thi 8th day of October 1974.

;(sEAL) fAttest:

McCOY M. GIBSON JR. c. MARSHALL DANN v :Attesting Officer Commissioner:of ,Patents

2. Composition as defined by claim 1 wherein said diol is selected fromthe group consisting of 1,3-octanediol, 1,3-heptanediol and1,3-nonanediol.
 3. Composition as defined by claim 1 wherein said diolester is selected from the group consisting of1,3-butanediol-1-monooctanoate and 1,3-octanediol-1-monopropionate. 4.Composition as defined by claim 1 wherein the amount of diol or diolester present is in the range from about 0.1 to 0.5 percent by weightbased upon the weight of the grain.
 5. Composition as defined by claim 1wherein said raw grain comprises wheat particles.
 6. Composition asdefined by claim 1 wherein said raw grain comprises oats.
 7. Compositionas defined by claim 1 wherein said raw grain comprises rye. 8.Composition as defined by claim 1 wherein said raw grain comprisesbeans.
 9. Composition as defined by claim 1 wherein said raw graincomprises maize.
 10. Composition as defined by claim 1 wherein said rawgrain comprises hay.
 11. Composition as defined by claim 1 wherein saidraw grain comprises rice.